CN108918500A - SERS method for separating based on immunomagnetic beads label - Google Patents
SERS method for separating based on immunomagnetic beads label Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000004416 surface enhanced Raman spectroscopy Methods 0.000 title claims abstract description 24
- 238000001514 detection method Methods 0.000 claims abstract description 34
- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 30
- 238000012360 testing method Methods 0.000 claims abstract description 6
- 238000012216 screening Methods 0.000 claims description 18
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B07—SEPARATING SOLIDS FROM SOLIDS; SORTING
- B07C—POSTAL SORTING; SORTING INDIVIDUAL ARTICLES, OR BULK MATERIAL FIT TO BE SORTED PIECE-MEAL, e.g. BY PICKING
- B07C5/00—Sorting according to a characteristic or feature of the articles or material being sorted, e.g. by control effected by devices which detect or measure such characteristic or feature; Sorting by manually actuated devices, e.g. switches
- B07C5/34—Sorting according to other particular properties
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G01N2015/1028—
Abstract
The present invention proposes a kind of SERS method for separating based on immunomagnetic beads label, including：S1. on the distinctive surface receptor of every kind of target cell, label is modified with the immunomagnetic beads with the antibody of surface receptor pairing；S2. target cell is filtered out；S3. the surface receptor according to specific to every kind of target cell, respectively on every kind of target cell, label is modified with the SERS-TAG with the antibody of surface receptor pairing；S4. surface-enhanced Raman detection is carried out to determine its type to each target cell after label respectively, is classified according to the type of target cell to target cell.The present invention can complete a variety of CTC detections in one test, and can greatly reduce the quantity of surface-enhanced Raman detection and the dosage of SERS-TAG, reduce the optical detection time, and save the cost.
Description
Technical field
The invention belongs to the interleaving techniques fields of cell sorting and optical engineering, and in particular to one kind is based on immunomagnetic beads mark
The SERS method for separating of note.
Background technique
Immunomagnetic beads have been widely applied in terms of molecule and cell biology, including nucleic acid extraction, specific protein
Sorting and enrichment of matter and cell etc..By taking the cell sorting based on immunomagnetic beads as an example, which is divided into four steps：(1)
Magnetic bead is surface modified with ligands specific or antibody；(2) by the magnetic bead by modification by antigen-antibody response or
Ligand-receptor association reaction is combined with target cell in sample；(3) pass through magnetic-adsorption magnetic bead-cell combination body；
(4) other cells are removed by means such as flushings.
The existing cell-based screening technology based on immunomagnetic beads directly passes through magnetic force and attracts magnetic bead, cause its for
The immunomagnetic beads of cell combination and unbonded immunomagnetic beads do not have separating capacity, are carrying out surface-enhanced Raman, miscellaneous to magnetic bead
The noise signal as caused by unbonded magnetic bead can not be removed by dissipating magnetic-field measurement etc. when further qualitative/quantitative detects, be limited
Immunomagnetic beads are further applied.
In laboratory research, international top research unit is by preparing specific antibody in Magnetic Sensor upper surface
Mode, and targeted cell surface antigen binding, to successfully capturing target cell above Magnetic Sensor.It is rushed again by fluid
It washes, so that it may realize the screening of Magnetic labeled cells.
But in the detection scheme of existing laboratory Magnetic Sensor, above targeted cell surface antigen and sensor
The combination of antibody removes unbonded magnetic bead by fluid scouring to realize the capture of target cell.The program can be removed
Other cells and unbonded magnetic bead are removed, but are difficult to ensure that micro target cell is successfully captured.Also, this scheme uses
Chemically combined mode is difficult to ensure the cell of the deniers such as circulating tumor cell and successfully captures, thus can be in diagnosis
Cause mistaken diagnosis.In addition, this method for capturing target cell by antibody-antigen binding needs artificial participate in and it is necessary to need
It wants technical professional that can complete, is unfavorable for integrating, be unable to satisfy the information system requirement of big data era.
In addition, antigen distinctive on target cell or receptor are marked using immunomagnetic beads, this method is specific
In the case of can only sub-elect a kind of target cell, can not confirm in whole blood sample whether to contain other type tumour cells.But
When in practical applications, such as to whole blood sample carrying out tumour cell detection, a kind of tumour cell is only isolated, is not enough to confirm
Whether also contain other kinds of tumour cell in whole blood sample, can not be further the detection of tumour cell (such as DNA sequencing
Deng) accurate detection sample message is provided.Meanwhile identical antigen or receptor are also likely to be present on different tumour cells, it uses
Kinds of tumor cells can be once sub-elected after immunomagnetic beads label, at this time, it is difficult to the specific type for determining tumour cell, it can not
Accurate detection sample message is provided for subsequent cancer diagnosis, and then saves subsequent detection period and cost.And it directly utilizes
SERS detection, is needed to be marked using a large amount of SERS-TAG, causes to waste, and very long by the optical system time, efficiency
It is extremely low.
Summary of the invention
The purpose of the present invention is to provide a kind of SERS method for separating based on immunomagnetic beads label, this method uses physics
Mode realize cell screening, greatly improve the capture rate of target cell, and a variety of CTC inspection can be completed in one test
It surveys, and the quantity of surface-enhanced Raman detection and the dosage of SERS-TAG can be greatly reduced, reduce the optical detection time and save
Cost.
To achieve the goals above, the present invention adopts the following technical scheme that：The sorting side SERS based on immunomagnetic beads label
Method includes the following steps：
A kind of SERS method for separating based on immunomagnetic beads label, includes the following steps：
S1. on the distinctive surface receptor of every kind of target cell, label is modified with the antibody with surface receptor pairing
Immunomagnetic beads；
S2. target cell is filtered out；
S3. the surface receptor according to specific to every kind of target cell, respectively on every kind of target cell, label be modified with
The SERS-TAG (surface-enhanced Raman label) of the antibody of surface receptor pairing；
S4. surface-enhanced Raman detection is carried out to determine its type, according to mesh to each target cell after label respectively
The type of mark cell classifies to target cell.
Further, the target cell after step S1 label is screened by cell sorting devices, it is described thin
The main structure of born of the same parents' screening plant is cut into upper layer, lower layer's cavity by one layer of cells screening, and cell buffering is filled in the cavity
Liquid；The concrete operation step of screening is：
It S21. will include free immunomagnetic beads, defeated by the sample of the target cell of immunomagnetic beads label and other cells
Enter into lower layer's cavity of cell sorting devices；
S22. longitudinal magnetic field and swaying are applied to cell sorting devices, immunomagnetic beads is sucked into upper layer cavity, target
Cell is adsorbed on the lower surface of cell sieve；
S23. it rinses respectively and obtains the solution comprising free immunomagnetic beads, the solution comprising other cells and comprising being exempted from
The solution of the target cell of epidemic disease marked by magnetic bead.
Further, the concrete operation step of the step S23 is：
S231. longitudinal magnetic field and swaying are cancelled, upper layer cavity is rinsed, is obtained comprising free immunomagnetic beads
Solution；
S232. apply longitudinal magnetic field, lower layer's cavity is rinsed, obtain the solution comprising other cells；
S233. longitudinal magnetic field is cancelled, lower layer's cavity is rinsed, obtains the target cell comprising being marked by immunomagnetic beads
Solution.
Further, the concrete operation step of the step S23 can also be：
S231. swaying is cancelled, lower layer's cavity is rinsed, obtains the solution comprising other cells；
S232. longitudinal magnetic field is cancelled, upper layer cavity is rinsed, obtains the solution comprising free immunomagnetic beads, it is right
Lower layer's cavity is rinsed, and obtains the solution of the target cell comprising being marked by immunomagnetic beads.
Further, the longitudinal magnetic field is generated by permanent magnet, soft magnetic bodies or electromagnet.
Further, the swaying includes swaying magnetic field and/or swaying wave and/or lateral water
Stream.
Further, the operation of the sample input and flushing is realized by syringe pump or miniflow pump.
Further, in step s 4, the target cell after step S3 label is classified by micro-fluidic system, it is described
Several output channels that micro-fluidic system includes input channel and is connected with input channel are provided with Raman inspection in input channel
The concrete operation step of measuring point, the step S4 is：
S41. target cell is made to form unicellular stream in input channel；
S42. surface-enhanced Raman detection is carried out to determine its type to the target cell by Raman detection point；
S43. according to the type of target cell, physics field force is applied to target cell, by different types of target cell point
From to being collected in different output channels.
Further, in the step S43, the physical field applied to cell includes electric field, magnetic field, light field and fluid field.
Further, the Raman detection point is provided with Raman spectrometer.
Compared with prior art, the advantages and positive effects of the present invention are：
(1) method of the invention passes through twice in targeted cell surface markers, and first time labelled immune magnetic bead can be simultaneously
All target cells are screened, different SERS-TAG is marked then can to complete to divide by type to target cell for the second time
Choosing.Method of the invention can detect plurality of target cell in one test, and they according to type be sorted out, energy
Accurate detection enough is provided for medical diagnosis on disease as a result, improving application of the immunomagnetic beads in clinical detection；
(2) present invention realizes cell screening using physics mode, by magnetic field gradient, irrigation flow rate, cavity size
Adjustment, realizes the high capture rate of target cell, will substantially increase the screening effect of target cell.Meanwhile free immunization magnetic bead
Sift out the detection of achievable high s/n ratio magnetic cell；
(3) method of the invention is easy to operate, and difficulty is small, moreover, the device for completing the method for the present invention is easily integrated
Sensor can satisfy the information system requirement of big data era.
Detailed description of the invention
Surface receptor schematic diagram of the Fig. 1 for first target cell and second target cell and thereon；
Fig. 2 is a kind of structural schematic diagram of cell sorting devices；
Fig. 3 is screening step operation chart；Wherein, (a) is the operation chart of step S21；(b) for step S22's
Operation chart；(c) operation chart for being step S231；(d) operation chart for being step S232；It (e) is step S233
Operation chart；
Fig. 4 is the cell microfluidic separation system schematic diagram based on SERS.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, right below in conjunction with drawings and examples
The present invention is further elaborated.It should be appreciated that described herein, specific examples are only used to explain the present invention, not
For limiting the present invention.
Embodiment 1
SERS method for separating based on immunomagnetic beads label of the invention, includes the following steps,
S1. on the distinctive surface receptor of every kind of target cell, label is modified with the antibody with surface receptor pairing
Immunomagnetic beads；
By taking whole blood cells as an example, it is assumed that the target cell for needing to detect has first target cell and two kinds of second target cell, choosing
When selecting surface receptor, specific surface receptor, as first target cell on first target cell and second target cell are selected
With the surface receptor all having on second target cell, and this surface receptor is no on other cells.Such as Fig. 1 institute
Show, be different from other cells, surface receptor specific to first target cell is a ', b ' and c ', table specific to second target cell
Face receptor is a ', b ' and d ', can be with then surface receptor a ' and b ' are that first target cell and second target cell are specific
One is selected to regard surface receptor, for example the antibody of selection surface receptor a ', surface receptor a ' pairing is a, then is modified with antibody a
Then immunomagnetic beads remove label first target cell and second target cell with the immunomagnetic beads after modification.
Wherein, the material of immunomagnetic beads can be the magnetic materials such as ferroso-ferric oxide, and magnetic bead outer layer is provided with coating, coated
Material can be the capsulating materials such as polystyrene, polyethyleneimine, polyvinyl alcohol, polyvinyl acetate, polysaccharide, in the present embodiment
In, magnetic bead is made of ferroso-ferric oxide, and diameter is 20 nanometers, and magnetic bead outer layer is coated with by polystyrene.
S2. target cell is filtered out；
Target cell after step S1 label is screened by cell sorting devices.Cell as shown in Figure 2
Screening plant, cell sorting devices include the rectangular-shape shell of inner hollow, and enclosure interior is horizontally disposed with one layer of cells sieve will
Shell is divided into upper layer cavity and lower layer's cavity, and phosphate buffer PBS is filled in cavity.The left side of cell sorting devices,
Pairing lower layer's cavity is provided with injection port；The right side of cell sorting devices, matches upper layer cavity and lower layer's cavity is respectively arranged with
Upper layer outlet and lower layer's outlet.
In the present embodiment, rectangular-shape is set by the shell of cell sorting devices, only a kind of preferred embodiment, when
So, the shell of cell sorting devices can be the regular shapes such as cylinder, square, can also be any irregular shape, ability
The technical staff in domain can be specifically arranged as needed.
Wherein, cell sieve can be made of silicon wafer, can also other can by dimethyl silicone polymer, polymethyl methacrylate etc.
Row material is made.For the material of cell sieve, shape, size and the arrangement of mesh are without rigid requirement, as long as the size of mesh is situated between
Between the diameter of target cell and the diameter of immunomagnetic beads, the generally circular or slit-type of mesh, net
Hole can be prepared by technologies such as nano impression or photoetching.In the present embodiment, 650 microns of thick silicon wafers of cell screening
It is prepared through photoetching technique.And mesh select slit-type, specifically use a length of 28 microns, wide 8 microns of quadratic mesh, mesh
Between spacing be 8 microns.
Specific step is as follows for screening：
It S21. will include free immunomagnetic beads, by the target cell of immunomagnetic beads label and its as shown in Fig. 3 (a)
The sample input of his cell is into lower layer's cavity of cell sorting devices.
S22. as shown in Fig. 3 (b), longitudinal magnetic field and swaying is applied to cell sorting devices, immunomagnetic beads are sucked
Upper layer cavity, target cell are adsorbed on the lower surface of cell sieve.
The effect of longitudinal magnetic field be immunomagnetic beads are attracted by magnetic field so that free immunomagnetic beads, by immunomagnetic beads
The target cell of label moves upwards, and longitudinal magnetic field is generally generated by permanent magnet, soft magnetic bodies or electromagnet.The work of swaying
With being the immunomagnetic beads weak vibrations so that free, so that immunomagnetic beads when by cell sieve, have high percent of pass.
Swaying can be magnetic field and generate swaying magnetic field, be also possible to a kind of swaying wave, including microwave, ultrasonic wave etc..
In the present embodiment, it selectes longitudinal magnetic field permanent magnet made of NdFeB material to generate, ndfeb magnet has excellent magnetic
Performance, the surface residual intensity of magnetization is up to 2000Oe.Swaying generates swaying magnetic field by electromagnet and provides, electromagnet knot
Structure is simple, is easy to control, and can adjust the size in magnetic field by adjusting the number of turns and coiling diameter of magnetic coil, usually shakes
Set of frequency is swung in 1-100HZ.In the present embodiment, 100Oe is selected, the oscillating magnetic gradient fields of 100HZ are used for magnetic bead
Apply lateral magnetic force, rate is sifted out in raising.
Before carrying out step S21, in the two sides of cell sorting devices, magnetic coil is respectively set, and be powered, in cell
Place ndfeb magnet in the side of screening plant upper layer cavity.
Apply magnetic field after, free immunomagnetic beads and by immunomagnetic beads mark target cell under the active force in magnetic field,
It is moved towards cell sieve, free immunomagnetic beads diameter is usually less than 500nm, much smaller than the mesh size of cell sieve, therefore
The target cell diameter that mesh can be passed through and enter upper layer cavity, and marked by immunomagnetic beads usually at 15 μm or more (such as
Tumour cell) it is greater than the mesh of cell sieve, therefore stopped by cell sieve.In the present embodiment, if cell sieve selects circle
Design, then the mesh of some cell sieve will be stopped completely by target cell, some possible free magnetic bead
It will remain in lower layer's cavity, and selecting quadratic mesh then is not in this problem, the target for the marked by magnetic bead being immunized
Although cell due to magnetic field effect and be adsorbed in cell sieve, will not be complete by the mesh of rectangle because shape is different
Obstruction, since the volume of immunomagnetic beads is much smaller for the target marked by immunomagnetic beads, immunomagnetic beads energy
Enough smoothly to pass through mesh, improve magnetic bead sifts out rate.
Further, the concrete operation step of the step S23 is：
S231. as shown in Fig. 3 (c), magnetic field longitudinal magnetic field and swaying is cancelled, upper layer cavity is rinsed, is obtained
Include the solution of free immunomagnetic beads, upper layer cavity be rinsed, obtain include free immunomagnetic beads solution.
Magnetic field longitudinal magnetic field and swaying are cancelled, the magnetic force that free immunomagnetic beads are subject to disappears, and passes through
To upper layer cavity rinse, can be easy to get only include free immunomagnetic beads solution.
S232. as shown in Fig. 3 (d), apply longitudinal magnetic field, lower layer's cavity is rinsed, obtains including other cells
Solution.
Apply longitudinal magnetic field, due to the suction-operated in magnetic field and the barrier effect of cell sieve, is marked by immunomagnetic beads
Target cell is firmly adsorbed on the downside of cell sieve, is selected suitable flow velocity to rinse lower layer's cavity at this time, can be obtained
To the solution for including other cells.
S233. as shown in Fig. 3 (e), longitudinal magnetic field is cancelled, lower layer's cavity is rinsed, obtained comprising by immunomagnetic beads
The solution of the target cell of label.
Longitudinal magnetic field is cancelled, the magnetic force being subject to by the target cell that immunomagnetic beads mark disappears, by lower layer
Cavity rinses, and can be obtained only includes by the solution of the target cell of immunomagnetic beads label.
The input of above-mentioned sample, and the flushing to upper and lower layer cavity are realized generally by pump, for example, when thin
When born of the same parents' screening plant is macro-scale devices, syringe pump can be used；When cell sorting devices are microdevices, miniflow can be used
Pump.The present embodiment controls the flowing of fluid sample by syringe pump, to obtain comprising different types of solution.
S3. the surface receptor according to specific to every kind of target cell, respectively on every kind of target cell, label be modified with
The SERS-TAG of the antibody of surface receptor pairing；
Surface receptor specific to every kind of target cell is selected to go to mark in step s3, it is thin with the target for distinguishing different
Born of the same parents, as shown in Figure 1, surface receptor specific to first target cell is b ' and c ', the antibody matched respectively is b and c；Second target is thin
Surface receptor specific to born of the same parents only has b ', and the antibody of surface receptor b ' pairing is b.Different SERS- is modified with antibody b and c
TAG forms two kinds of modifiers of b-SERS-TAG1 and c-SERS-TAG2, then goes label target thin with the SERS-TAG after modification
Born of the same parents, the label of two kinds of target cells is different at this time, and first target cell has the label of SERS-TAG1 and SERS-TAG2, and
Second target cell only has SERS-TAG1 label, two kinds of cell differentiations can thus be come by SERS spectrogram.
S4. surface-enhanced Raman detection is carried out to determine its type, according to mesh to each target cell after label respectively
The type of mark cell classifies to target cell.With specific reference to Fig. 4.
The concrete operation step of step S4 is：
S41. target cell is made to form unicellular stream in input channel；
The concrete methods of realizing of step S41 has, and target cell is input into input channel in the form of unicellular stream, or
Input channel is sized to only to allow unicellular pass through.Although input channel is sized to only allow unicellular
By that also input sample can be made to form unicellular stream in input channel, but in actual operation, the inlet of input channel
It is easy to be adhered cell mass together and block, therefore use in the present embodiment input sample is defeated in the form of unicellular stream
Into in input channel.
S42. surface-enhanced Raman detection is carried out to determine its type to the target cell by Raman detection point；
Raman spectrometer is provided on Raman detection point, Raman spectrometer receives Raman for emitting Raman spectrum
Scattering spectrum.
Input sample forms unicellular stream in input channel, when single target cell passes through Raman detection point, Raman
On spectral illumination to the target cell, it may occur that Raman scattering is analyzed the raman scattering spectrum detected, by wherein
The size and location of Raman peaks determines the type of cell.
S43. according to the type of target cell, physics field force is applied to target cell, by different types of target cell point
From to being collected in different output channels.
In order to by different types of target cell categorised collection, therefore an input channel is set to match several outputs logical
Road applies physics field force to target cell according to the type of the target cell detected, drive different types of target cell with
Into in different output channels.
Wherein, the physical field applied to cell includes electric field, magnetic field, light field and fluid field, can pass through adjusting physical field
The size and Orientation of power drives different types of cell to enter in the output channel of pairing.
The present embodiment is first elected a variety of CTC rough segmentations by magnetic bead, then is segmented the type of CTC by SERS.
First with magnetic bead, a variety of CTC are screened from a large amount of haemocyte, therefore greatly reduced with the SERS cell concentration being incubated for,
To reduce SERS dosage, also there is no need to whole blood cells all to pass through optical system.Chemistry sieve compared with the existing technology
Mode is selected, which realizes cell screening using physics mode, by the adjustment to magnetic field gradient, irrigation flow rate, cavity size,
The screening effect of micro target cell will be substantially increased.Meanwhile free immunization magnetic bead sifts out, it can be achieved that high s/n ratio is magnetic
Cell detection.And method of the invention can detect plurality of target cell in one test, and according to type by them
It sorts out, accurate detection can be provided for medical diagnosis on disease as a result, improving application of the immunomagnetic beads in clinical detection.Together
When the test sample that clearly classify can be provided for succeeding target cell detection (such as DNA detection), and then improve subsequent detection
Efficiency.
Embodiment 2
Embodiment 2 difference from example 1 is that, in the present embodiment swaying select swaying microwave, by
Microwave generator generates.Immunomagnetic beads are under the active force of horizontal miniwave, weak vibrations, rate of the immunomagnetic beads Jing Guo cell sieve
It is all very high with percent of pass.
Embodiment 3
Embodiment 3 difference from example 1 is that, in the present embodiment swaying select lateral water flow.Exempt from
Epidemic disease magnetic bead is under the active force of lateral water flow, weak vibrations, and rate and percent of pass of the immunomagnetic beads Jing Guo cell sieve are all very
It is high.
Embodiment 4
Embodiment 4 and implement 1 the difference is that, in the present embodiment, step S23 is specially：
Step S231 cancels swaying, is rinsed to lower layer's cavity, obtains the solution comprising other cells；
Step S232 cancels longitudinal magnetic field, is rinsed to upper layer cavity, obtains comprising the molten of free immunomagnetic beads
Liquid is rinsed lower layer's cavity, obtains the solution of the target cell comprising being marked by immunomagnetic beads.
Operating procedure in embodiment 4 is easier.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
It imitates embodiment and is applied to other fields, but without departing from the technical solutions of the present invention, according to the technical essence of the invention
Any simple modification, equivalent variations and remodeling to the above embodiments, still fall within the protection scope of technical solution of the present invention.
Claims (10)
1. the SERS method for separating based on immunomagnetic beads label, which is characterized in that include the following steps,
S1. on the distinctive surface receptor of every kind of target cell, label is modified with exempting from for the antibody matched with the surface receptor
Epidemic disease magnetic bead；
S2. target cell is filtered out；
S3. the surface receptor according to specific to every kind of target cell, respectively on every kind of target cell, label is modified with and the table
The SERS-TAG (surface-enhanced Raman label) of the antibody of face acceptor pair；
S4. surface-enhanced Raman is carried out to each target cell after label respectively to detect to determine its type, it is thin according to target
The type of born of the same parents classifies to target cell.
2. the SERS method for separating according to claim 1 based on immunomagnetic beads label, which is characterized in that step will be passed through
Target cell after S1 label is screened by cell sorting devices, and the main structure of the cell sorting devices is thin by one layer
Born of the same parents' screening is cut into upper layer, lower layer's cavity, is filled with Cell Buffer in the cavity；The concrete operation step of screening is：
S21. it will include free immunomagnetic beads, be arrived by target cell and the sample inputs of other cells that immunomagnetic beads mark
In lower layer's cavity of cell sorting devices；
S22. longitudinal magnetic field and swaying are applied to cell sorting devices, immunomagnetic beads is sucked into upper layer cavity, target cell
It is adsorbed on the lower surface of cell sieve；
S23. it rinses respectively and obtains the solution comprising free immunomagnetic beads, the solution comprising other cells and comprising by immune magnetic
The solution of the target cell of pearl label.
3. the SERS method for separating according to claim 2 based on immunomagnetic beads label, which is characterized in that the step
The concrete operation step of S23 is：
S231. longitudinal magnetic field and swaying are cancelled, upper layer cavity is rinsed, is obtained comprising the molten of free immunomagnetic beads
Liquid；
S232. apply longitudinal magnetic field, lower layer's cavity is rinsed, obtain the solution comprising other cells；
S233. longitudinal magnetic field is cancelled, lower layer's cavity is rinsed, obtains including the molten of the target cell marked by immunomagnetic beads
Liquid.
4. the SERS method for separating according to claim 2 based on immunomagnetic beads label, which is characterized in that the step
The concrete operation step of S23 is：
S231. swaying is cancelled, lower layer's cavity is rinsed, obtains the solution comprising other cells；
S232. longitudinal magnetic field is cancelled, upper layer cavity is rinsed, the solution comprising free immunomagnetic beads is obtained, to lower layer
Cavity is rinsed, and obtains the solution of the target cell comprising being marked by immunomagnetic beads.
5. the SERS method for separating based on immunomagnetic beads label according to Claims 2 or 3 or 4, it is characterised in that：It is described
Longitudinal magnetic field is generated by permanent magnet, soft magnetic bodies or electromagnet.
6. the SERS method for separating based on immunomagnetic beads label according to Claims 2 or 3 or 4, it is characterised in that：It is described
Swaying includes swaying magnetic field and/or swaying wave and/or lateral water flow.
7. the SERS method for separating based on immunomagnetic beads label according to Claims 2 or 3 or 4, it is characterised in that：It is described
The operation of sample input and flushing is realized by syringe pump or miniflow pump.
8. the SERS method for separating according to claim 1 based on immunomagnetic beads label, which is characterized in that in step S4
In, the target cell after step S3 label is classified by micro-fluidic system, the micro-fluidic system include input channel and
Several output channels being connected with input channel are provided with Raman detection point, the specific behaviour of the step S4 in input channel
It is as step：
S41. target cell is made to form unicellular stream in input channel；
S42. surface-enhanced Raman detection is carried out to determine its type to the target cell by Raman detection point；
S43. according to the type of target cell, physics field force is applied to target cell, by different types of target cell separate to
It is collected in different output channels.
9. the SERS method for separating according to claim 8 based on immunomagnetic beads label, which is characterized in that the step
In S43, the physical field applied to cell includes electric field, magnetic field, light field and fluid field.
10. the SERS method for separating according to claim 8 based on immunomagnetic beads label, which is characterized in that the Raman
Test point is provided with Raman spectrometer.
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